Flow Cytometry Equipment

The Flow Cytometry Core Facility is equipped with the following:

  • Becton Dickinson FACSVantage SE Turbo Sorter with DIVA Option . This instrument is capable of high speed and single cell sorting on up to ten parameters (forward & side scatter and 8 fluorescence detectors). The instrument is equipped with three lasers: 488nm air cooled, 633 nm HeNe, and multi-line water cooled laser with UV capability. Pulse Processing capabilities allow the use of area and width signals for doublet discrimination or parameter ratioing for the evaluation of Calcium Flux using Indo-1. The instrument has the capability of four way sorting into tubes or directly into plates (6-96 wells) or slides. It is equipped with an Aerosol Containment System for the sorting of BSL-2 biohazardous samples. Examples of possible fluorochromes which can be evaluated with this instrument:
    • 488nm excitable: FITC, PE, PE-Cy5, PE-Cy5.5, PE-Cy7, PE-Texas Red, Cy3, GFP, YFP, DsRed, CFSE, Alexa 488, PI, 7-AAD, Acridine Orange, PKH-26, DiI, DHR123, Fura Red, Fluo 3, DCFDA, Sytox Green, DRAQ 5, Ethidium Bromide, JC-1, TMR
    • 633nm excitable: APC, APC-Cy5.5, APC-Cy7, Cy5, Alexa 633, Alexa 647, TO-PRO3, DRAQ 5
    • UV (351nm) excitable: Hoechst 33342, Hoechst 33258, DAPI, Alexa 350, EBFP, Indo-1, Fluorogold
      FACSVANTAGE DIVA
    Consult the facility for available filter set ups, fluorochrome combinations, and appropriate controls, prior to designing your experiment. Located in Jordan Hall, Room 7065; 434-243-2695. See Scheduling for information on scheduling time on this instrument.

 

  • Two Becton Dickinson FACSCalibur dual laser (488nm and 635nm) benchtop cytometers (FACSCalibur B and C) capable of 7 parameter (four color) analysis. These instruments  can detect three fluorochromes excited by the 488nm laser and one fluorochrome excited by the 635nm laser.
  • One modified Becton Dickinson FACSCalibur dual laser (488nm and 637nm) benchtop cytometer (FACSCalibur A) capable of 7 parameter (five color) analysis.   This instrument  can detect three fluorochromes excited by the 488nm laser and two fluorochromes excited by the 637nm laser.
  • Examples of possible fluorochromes which can be evaluated on these instruments:
    • 488nm excitable: FITC, PE, PerCP, PE-Cy5, PE-Cy5.5, PE-Cy7, PerCP-Cy5.5, CFSE, GFP, Alexa 488, PI, 7-AAD, DRAQ 5, Acridine Orange, PKH-26, Fura Red, Fluo-3, DiI, DCFDA, Sytox Green, Cy3, Ethidium Bromide, DHR123, TMR, JC-1
    • 635 excitable (dual laser instrument only): APC, DRAQ 5, TO-PRO3, Cy5, Alexa 633, Alexa 647, APC-CY7 (FACSCalibur A only), and APC-Cy5.5 (FACSCalibur A only)
    Emission filters on this instrument are fixed and are as follows:
    • Emissions from the 488nm excitation
      • FL1 - 530/30 nm (515-545 nm)
      • FL2 - 585/42 nm (564-606 nm)
      • FL3 - 670 nm LP (>670 nm)
    • Emissions from the 635 nm excitation (FACSCalibur B &C)
      • FL4 - 661/16 nm (653-669 nm)
    • Emissions from the 637nm excitation (FACSCalibur A)
      • FL4 - 667/27
      • FL5 - 765/20

    Consult the facility for fluorochrome combination feasibility and necessary controls prior to designing your experiment. Located in Jordan Hall, Room 7051; 434-243-2711.

    All of these instruments can be run with a 96 well plate automated sample acquisition system.

    • See Scheduling for information on scheduling time for these instruments.

amsgif  FACS Calibur

  • CyAn™ ADP LX 9 Color
    CyAn™ ADP 9 Color has 11 parameters (FSC, SCC, FL1-FL9) and is equipped with 488 nm, 633 nm, and 405 nm solid state lasers. This model is ideal for up to 9 color analysis. Typical fluorochromes which can be used are FITC, GFP, CFSE, Alexa488, PE, PE-Cy5, PI, PE-Cy5.5,PER-CP, 7AAD, PE-Cy7, APC, Alexa647, Alexa633, APC-Cy7, APC-Cy5.5, Pacific Blue and Cascade Yellow, Pacific Orange, Cyan Fluorescent Protein.
    Click here to view optical layout

                      Cyan ADP
Consult the facility for fluorochrome combination feasibility and necessary controls prior to designing your experiment. Located in Jordan Hall, Room 7051; 434-243-2711.

 

  • Amnis Imagestream100TM
    Imagestream100
    An Entirely New Way to Analyze Cells
    By combining microscopy and flow cytometry in a single platform, the ImageStream  achieves a true breakthrough in cell analysis. Now the visual information that you obtain with a microscope can be fully integrated with the population statistics you would typically get from a flow cytometer. You can define and quantify cell subpopulations not only by signal intensity, but by the location of the signal in or on the cell. You can use cell and nuclear morphologies to characterize cell populations. In short, you can take full advantage of complex visual data to perform robust, multiparameter quantitative analyses.
    In the ImageStream , cells are illuminated with both a brightfield source and  405nm, 488nm and 658nm lasers. The resulting composite images are spectrally decomposed into as many as 6 component images and captured using Time Delay Integration technology on a proprietary CCD camera. You see a brightfield, darkfield and up to 4 fluorescence images of every cell, while over 30 morphological and signal intensity features are calculated for every image - nearly 200 features in total. With the new Extended Depth of Field option the ImageStream depth of field to be is expanded more than 5-fold, enabling such applications as FISH In Suspension (FISHIS®), as well as many other resolution-sensitive applications.
    High resolution images deliver quantitative information about cell structure and about the location of your target molecules. Follow translocation from the cytoplasm to the nucleus; quantify capping and co-capping reactions; separate cells based on unique morphologic criteria; study cell-cell interactions; differentiate cell states such as late apoptosis and necrosis -- the range of potential applications is extremely broad.
    For more details on sample preparation and fluorochromes choices click here.
  • Luminex 100 ISTM System
    Luminex IS100
    A flexible analyzer based on the principles of flow cytometry enables you to multiplex (simultaneously measure) up to 100 analytes in a single microplate well, using very small sample volumes. The system delivers fast and cost-effective bioassay results on many assay formats including nucleic acid assays, receptor-ligand assays, immunoassays and enzymatic assays.
    • For a list of companies who supply reagent kits for use on the Luminex platform click here.
    • For a list of applications for this technology click here.

  • Data Analysis Workstations
    The facility makes available both PC and Mac data analysis workstations. These workstations have a variety of data analysis software packages which can be used to analyze immunophenotyping, DNA, image, and multiplex analyte data. These workstations can be scheduled in the same manner as scheduling an instrument using Corporate Time.
    • Mac G5 workstation (1) - single processor, 1 GB RAM, CD burner, USB compatible. Software: FlowJo, Modfit, CelquestPro, Microsoft Office Suite, Toast 5.0.
    • Mac G5 workstation (2) - dual processor, 3 GB RAM, CD/DVD burner, USB compatible. Software: FlowJo, Modfit, CelquestPro, Microsoft Office Suite, Toast 5.0
    • PC Workstation - Pentium 4, 3GHz, 1 GB RAM, 16X R/W DVD. Software: IDEASTM Statistical Image Analysis Software, BeadviewTM  Multiplex Data Analysis Software, Coming Soon! -FCS ExpressTM Flow Cytometry Data Analysis Software, SummitTM Flow Cytometry Data Analysis Software.